LOCUS       V028197                 8106 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028197
VERSION     V028197
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2786
                     /label="DPYSL2(NM_001197293)"
                     /note="DPYSL2(NM_001197293)"
                     /gene="DPYSL2"
     CDS             2793..2858
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2882..3470
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3536..3584
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3786..4214
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4228..4557
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4624..5418
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5594..5715
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5764..5780)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5788..5804
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5812..5842)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5857..5878
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6166..6754)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6925..7785)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7786..7890)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc cgagagaaag caatccggga
      781 aggcggcaga ggacgaagag gtccctgctt tttttaaaaa cctgggctcc ggcagcccca
      841 agccccggca gaaattctgt ggcatgttct gcccggtgga agggtcctcg gagaacaaga
      901 ccatcgactt cgactcgctg tcggtgggcc ggggctcggg gcaggtggtg gctcagcagc
      961 gggacgtcgc ccacttgggc ccggacccgc agccgccgta ctcgcggcag ggccggcgcg
     1021 ccggcggaga gccatctgtt gaatcgggcc ggaaggtgga gatccggagg gcctcgggca
     1081 aagaagccct gcagaacatc aacgaccaga gcgatcgtct tctgatcaaa ggaggtaaaa
     1141 ttgttaatga tgaccagtcg ttctatgcag acatatacat ggaagatggg ttgatcaagc
     1201 aaataggaga aaatctgatt gtgccaggag gagtgaagac catcgaggcc cactcccgga
     1261 tggtgatccc cggaggaatt gacgtccaca ctcgtttcca gatgcctgat cagggaatga
     1321 cgtctgctga tgatttcttc caaggaacca aggcggccct ggctggggga accactatga
     1381 tcattgacca cgttgttcct gagcctggga caagcctgct cgctgccttt gaccagtgga
     1441 gggaatgggc cgacagcaag tcctgctgtg actactctct gcatgtggac atcagtgagt
     1501 ggcataaggg catccaggag gagatggaag cgcttgtgaa ggatcacggg gtaaattcct
     1561 tcctcgtgta catggctttc aaagatcgct tccagctaac ggattgccag atttatgaag
     1621 tactgagtgt gatccgggat attggcgcca tagcccaagt ccacgcagaa aatggcgaca
     1681 tcattgcaga ggagcagcag aggatcctgg atctgggcat cacgggcccc gagggacatg
     1741 tgctgagccg acctgaggag gtcgaggccg aagccgtgaa tcgtgccatc accatcgcca
     1801 accagaccaa ctgcccgctg tatatcacca aggtgatgag caaaagctct gctgaggtca
     1861 tcgcccaggc acggaagaag ggaactgtgg tgtatggcga gcccatcact gccagcttgg
     1921 gaacggacgg ctcccattac tggagcaaga actgggccaa ggctgctgcc tttgtcacct
     1981 ccccaccctt gagccctgat ccaaccactc cagactttct caactccttg ctgtcctgtg
     2041 gagacctcca ggtcacgggc agtgcccatt gcacgtttaa cactgcccag aaggctgtag
     2101 gaaaggacaa cttcaccctg attccggagg gcaccaatgg cactgaggag cggatgtccg
     2161 tcatctggga caaggctgtg gtcactggga agatggatga gaaccagttt gtggctgtga
     2221 ccagcaccaa tgcagccaaa gtcttcaacc tttacccccg gaaaggccgc attgctgtgg
     2281 gatccgatgc cgacctggtc atctgggacc ccgacagcgt taaaaccatc tctgccaaga
     2341 cacacaacag ctctctcgag tacaacatct ttgaaggcat ggagtgccgc ggctccccac
     2401 tggtggtcat cagccagggg aagattgtcc tggaggacgg caccctgcat gtcaccgaag
     2461 gctctggacg ctacattccc cggaagccct tccctgattt tgtttacaag cgtatcaagg
     2521 caaggagcag gctggctgag ctgagagggg ttcctcgtgg cctgtatgac ggacctgtgt
     2581 gtgaagtgtc tgtgacgccc aagacagtca ctccagcctc ctcggccaag acgtctcctg
     2641 ccaagcagca ggccccacct gtccggaacc tgcaccagtc tggattcagt ttgtctggtg
     2701 ctcagattga tgacaacatt ccccgccgca ccacccagcg tatcgtggcg ccccccggtg
     2761 gccgtgccaa catcaccagc ctgggcggat ccgactacaa agaccatgac ggtgattata
     2821 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga
     2881 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2941 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     3001 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     3061 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     3121 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     3181 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     3241 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     3301 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     3361 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     3421 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg
     3481 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     3541 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     3601 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     3661 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     3721 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta
     3781 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3841 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3901 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     3961 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4021 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4081 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4141 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4201 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4261 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4321 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4381 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4441 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4501 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4561 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4621 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4681 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4741 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4801 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4861 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4921 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4981 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5041 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5101 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     5161 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5221 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5281 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5341 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5401 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     5461 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5521 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5581 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5641 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5701 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5761 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5821 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5881 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5941 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6001 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6061 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6121 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6181 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6241 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6301 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6361 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6421 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6481 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6541 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6601 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6661 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6721 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6781 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6841 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6901 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6961 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7021 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7081 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7141 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7201 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7261 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7321 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7381 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7441 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7501 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7561 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7621 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7681 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7741 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7801 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7861 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7921 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat
     7981 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     8041 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     8101 cttagg
//
